antidiabetic activity of hibiscus syriacus on alloxan ...antidiabetic activity of hibiscus syriacus...

VOLUME 4 | NUMBER 5 | OCT | 2014 |140

Available online at www.ijntps.org | ISSN: 2277 – 2782 INTERNATIONAL JOURNAL OF NOVEL TRENDS IN PHARMACEUTICAL SCIENCES

Antidiabetic activity of Hibiscus syriacus on alloxan induced

diabetic rats

Kanjeri Nirosha*1, M.Divya1, R. Himabindu1, N. Himaja1, M. Pooja1, Mohemmed Sadiq2

*1Sree Vidyanikethan College of Pharmacy, Sree Sainath Nagar, Tirupati, A.Ranganpet, Andhra Pradesh-517102. 2Krishna Teja Pharmacy College, Chadalawada Nagar, Renigunta Road, Renigunta Road, Tirupati – 517501.

INTRODUCTION

Diabetes mellitus is a group of metabolic diseases

characterized by elevated blood glucose levels. It is

a result of defects in insulin secretion, insulin action

or both. Non‑Insulin Dependent Diabetes Mellitus

(NIDDM) is a multifactorial disease characterised by

hyperglycemia and lipoprotein abnormalities. These

traits are hypoyhetized to damage cell membranes,

which results in excessive generation of reactive

oxygen species. NIDDM has also been associated

with an increased risk for developing premature

atherosclerosis due to an increase in Triglyceride

(TG) and Low Density Lipoproteins (LDL) and

decrease in High Density Lipoprotein (HDL) levels

[1]. Chronic hyperglycemia is associated with

microvascular and macrovascular complications that

can lead to visual impairment, blindness, kidney

disease, nerve damage, amputations, heart disease,

and stroke. In 1997 an estimated 4.5% of the US

population had diabetes. Direct and indirect health

care expenses were estimated at $98 billion [2].

Epidemiology

It is estimated that 366 million people had DM in

2011; by 2030 this would have risen to 552 million

[3] the number of people with type 2 DM is

increasing in every country with 80% of people with

DM living in low- and middle-income countries. DM

caused 4.6 million deaths in 2011. It is estimated

that 439 million people would have type 2 DM by

the year 2030 [4]. The incidence of type 2 DM varies

substantially from one geographical region to the

other as a result of environmental and lifestyle risk

factors [5].

MATERIALS AND METHOD

Plant material

The whole plant of Hibiscus syriacus was collected

in kanipakam, chittoor district. The plant material

was authentified by Dr.K.Madhava chetty,

Assistant professor, department of botany, S.V.

University, Tirupathi. The plaant material was dried

in shade at room temperature. Then dried material

was powdered with the help of mechanical grinder

and subjected to seiving for equal particles of

coarse powder with the help of seiver.

Preparation of extract

The powdered plant material was packed in to

Soxhlet apparatus and defatted with petroleum

ether (60 - 80°C). Marc left was further extracted

To whom correspondence should be addressed:

K. Nirosha

Email: [email protected]

Article Info

Article history

Received 05 Oct 2014

Revised 10 Oct 2014

Accepted 12Oct 2014

Available online 30Oct 2014

Keywords

Diabetes Mellitus, Hibiscus

Syriacus, Blood Glucose.

Abstract

Diabetes mellitus is characterized by elevated blood glucose levels. The present

study was to evaluate the antidiabetic potential of Hibiscus syriacus on alloxan

induced diabetic rats. Animals are divided into five groups. For group I was

administered with distilled water (10 ml/kg/p.o), group II with alloxan

(120mg/kg/i.p), group III administered with alloxan and metformin (150mg/kg/p.o),

group IV was received alloxan and ethanolic extract (200 mg/kg/p.o), group V

received alloxan and ethanolic extract (400mg/kg/p.o). Administration of test and

standard drug for the period of 28 days, on 28th day biochemical parameters were

estimated. Results shows the decreased blood glucose level. Antidiabetic potential

was high in group V compared to group IV. Hence It concluded that high dose of

ethanolic extract shows the more antidiabetic potential compared to low dose.

RESEARCH ARTICLE

K. Nirosha et al., Antidiabetic activity of Hibiscus syriacus on alloxan induced diabetic rats


with ethanol. Extract was air dried until it comes to

semi-solid to solid mass.

Phytochemical screening

The extract obtained was subjected to qualitative

tests for identification of different constituents like

tannins, alkaloids, saponins, glycosides, terpenes,

phenolics, flavonoids, carbohydrates, proteins and

steroids, by using simple and standard qualitative

methods described by Trease and Evans [6].

PHARMACOLOGICAL STUDY

Animals

Healthy albino rats of either sex weighing about

120-150 g were used during the study. The animals

were procured from Sree Vidyanikethan College of

Pharmacy, Tirupati, India. Before initiation of

experiment, the rats were acclimatized for a period

of 7 days. Standard environmental conditions such

as temperature ranging from 18 to 320C, relative

humidity ( 70%) and 12 hrs dark/light cycle were

maintained in the quarantine. All the animals were

fed with rodent pellet diet and water under strict

hygienic conditions. All procedures were performed

in accordance to CPCSEA guidelines after approval

from Sree Vidyanikethan College of Pharmacy,

Tirupati, India [Approval no. SVCP/IAEC/I-010/2013-

2014, dated on 14 march 2014.].

Acute oral toxicity study

The acute toxicity of ethanolic extract of Hibiscus

syriacus was carried out as per OECD-423 guidelines

for safe dose administration to animals. . The

animals were kept fasting for overnight providing

only water, after which the extracts were

administered orally at the dose level 2000mg/kg

body weight and observed for 14 days. the results

showed no sign of toxicity till 14 days at a starting

dose of 2000 mg/kg body weight. Hence, 1/10th

were considered as test dose for the study.

Induction of diabetes

Alloxan monohydrate was obtained from BROS

scientifics, tirupathi. To induce diabetes, alloxan 120

mg/kg, intraperitoneal (i.p.) Single dose will be

administered intraperitonial to groups II, III,IV&V.

The fasting blood glucose was determined after 72

hours7 Rats showing a blood glucose level of >200

mg/dl were taken for the study.

Experimental design

Rats having blood glucose level more than

200mg/dl taken for the study. Then rats are divided

into 5 groups, each group having 6 animals. Group

I: received normal saline (10ml/kg/day/p.o)

Group II: received alloxan (120mg/kg/i.p), Group III:

received standard drug Metformin (150mg/kg/p.o)

+ alloxan (120mg/kg/i.p), Group IV: received

ethanolic extract of Hibiscus syriacus

(200mg/kg/p.o) + alloxan (120mg/kg/i.p.), Group V:

received ethanolic extract of Hibiscus syriacus

(400mg/kg/p.o) + alloxan (120mg/kg/i.p.).

The study performed for 28 days.

Sample collection

28th day animals were sacrificed, pancreas was

isolated and it were fixed in 10% neutral formalin

solution. Fine sections were mounted on glass slides

and stained with hematoxylin Eosin for light

microscopic analysis.

Blood glucose estimation

Determination of blood glucose Fasting blood

glucose was estimated by commercially available

glucose strips (Accu- Chek) using One Touch

Glucometer (Johnson – Johnson, India). Estimation

of blood glucose level on 0th day, 7th day, 14th and

28thday.

RESULTS

Phytochemical screening: phytochemical screening of ethanolic extract was showed the presence of flavonoids,

tannins, saponins, glycosides and phenols. Carbohydrates, proteins, aminoacids and alkaloids are absent.

Table 1. Estimation of blood glucose level(mg/dl)

GROUPS 0TH DAY 7

TH DAY 14

TH DAY 28

TH DAY

Control 99.17±0.87 100±0.91 99.8±0.87 99.33±0.76

Diabetic control 230±2.9*a 230.5±2.91*a 235±1.68*a 239.7±0.98*a

Standard 230±3.1**b 157.7±1.9**b 116±1.40**b 113.2±0.90**b

EEHS(200mg/kg) 232±3.1*b 176±1.24*b 158±1.16*b 147.5±0.76*b

EEHS(400mg/kg) 232±3.1**b 162±1.72**b 129±2.62**b 128.2±o.70**b



Values are expressed in ±SEM values (n=6). One‑way ANOVA followed by Dunnet’s multiple comparison tests.

Alloxan (120mg/kg.) was injected to except control , all other drug groups; a alloxan induced diabetic group vs

normal group, *p <0.001; b extract treated group vs alloxan induced diabetic group, *p <0.01; **p<0.001.

Fig 1. Estimation of blood glucose level

Fig 2. Histopathology of pancreas

A B

C



D E

A – control- healthy β cells , B-Negative control- destruction of β cells, C-standard-recovery of β cells, D-

EEHS(200mg/kg)-slow recovery of β cells, E- EEHS(400mg/kg)-recovery of β cells better than “D”.

DISCUSSION

Pancreas is the primary organ involved in sensing

the organism’s dietary and energetic states via

glucose concentration in the blood and in response

to elevated blood glucose, insulin is Secreted.

Alloxan is one of the usual substances used for the

induction of diabetes mellitus apart from

streptozotocin by induce free radical production

and cause tissue injury. It has a destructive effect of

the beta cells of the pancreas [8]. A massive

reduction in insulin release by the destruction of

beta-cells of the islets of langerhans thereby

inducing hyperglycemia [9]. Alloxan has been

shown to Ethanolic Extract in the dose of 400

mg/kg reduced the reduction in the blood glucose

levels compared to Extract in the dose of 200

mg/kg. The literature reports reveal that flavonoids

and tannins present in the plant extract known to

possess antihyperglycemic activity.

Antihyperglycemic potential of test extract was

observed may be due to presence of similar

phytoconstituents which was evident by preliminary

phytochemical screening. In the present

investigation demonstrated ethanolic extract of

Hibiscus syriacus has the significant anti-diabetic

Activity.

CONLUSION

Present study was concluded that the ethanolic

extract of Hibiscus syriacus at the dose of 400 mg/kg

body weight produced significant antidiabetic

activity in alloxan-induced NIDDM in albino rats.

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